抗萘单克隆抗体杂交瘤细胞株的构建

为了构建针对石油运输泄露导致的海洋环境中萘等多环芳烃污染的免疫学快速检测方法，需获得高效稳定的分泌抗萘单克隆抗体的杂交瘤细胞株。本文以2-萘丁酸-KLH为免疫抗原免疫BALb/c小鼠，间接竞争ELISA测定小鼠血清效价，用细胞融合筛选技术筛选抗萘单克隆抗体杂交瘤细胞株，捕获ELISA测定小鼠免疫球蛋白亚型。结果显示，实验小鼠免疫血清效价最高能达到1:80000，经过HAT筛选、复检和亚克隆后得到5个能分泌抗萘单克隆抗体的细胞株，细胞株上清液效价最高能达到1:15625，抗萘单克隆抗体免疫球蛋白亚型均为IgG1，Kappa轻链。本方法成功构建的杂交瘤细胞株能稳定高效分泌抗萘单克隆抗体，抗体亚型符合免疫学检测的特性，为萘的ELISA检测方法建立及胶体金免疫层析（GICA）试纸条的研制提供了稳定高效的单克隆抗体源。

Development of hybridoma cell lines secreting monoclonal antibodies against naphthalene

In order to establish a rapid immunological detection method of naphthalene and other polycyclic aromatic hydrocarbons (PAHs), which originated from leakage of petroleum transportation in the marine, highly efficient and stable hybridoma cell line secreting anti-naphthalene McAb is urgently needed.In this study, 2-naphthyl butyric acid -KLH was used as immune antigen to immunize BALb/c mice in order to detect serum titer in mice coupled with indirect competitive ELISA. The technique of cell hybridoma screening was used to screen hybridoma cell line resistant to anti-naphthalene monoclonal antibody and determined the immunoglobulin subtype of mice with capture ELISA.Results showed that the mice serum titer can reach the maximum value of 1:80000.The five cell lines which can secrete anti-naphthalene monoclonal antibody after selection by HAT, and subclones whose cell supernatant titer can reach a maximum value of 1:15625.All of the immunoglobulin subtypes of anti\aphthalene monoclonal antibody were IgG1, Kappa light chain.As a result, the successfully constructed hybridoma cell lines can stably secrete anti-naphthalene monoclonal antibody, whose antibody subtypes agree with the immunological detection characteristics.Our method provides as a stable and efficient source of monoclonal antibody for establishing the detecting method of ELISA as well as a reference for developing the colloidal gold immunochromatographic method for ELISA detection of naphthalene (GICA) strip.